BIOAVAILABILITY

• The extent and rate at which its active moiety is

delivered from pharmaceutical form and becomes
available in the systemic circulation

Pharmacokinetics
conc. vs time
Conc.(mg/L)

0.0
0 25

Time (h)
 Definition:
• Bioavailability is defined as the fraction of unchanged drug reaching
the systemic circulation following administration by any route.

• To exert an optimal therapeutic action an active moiety should be

delivered to its site of action in an effective concentration for the
desired period.

 The bioavailability of an oral dosage form is determined by

comparing the Area Under Curve (AUC) after oral
administration of a single dose with that obtained when given IV

• Drug bioavailability = AUC (oral)

AUC (IV)

= Bioavailable dose
Administered dose
THE NEED FOR BIOAVAILABILITY STUDIES

Bioavailability studies provide and estimate of the fraction of the orally

administered dose that is absorbed into the systemic circulation when compared to
the bioavailability for a solution, suspension, or intravenous dosage form that is
completely available.

Bioavailability studies provide other useful information that is important to

establish dosage regimen and to support drug labeling, such as distribution and
elimination characteristics of the drug
Bioavailability studies provide information regarding the performance of the
formulation
 Route Bioavailability (%) Characteristics

Intravenous 100 (by definition) Most rapid onset

(IV)

Intramuscular 75 to ≤ 100 Large volumes often feasible; may be

(IM) painful

Subcutaneous 75 to ≤ 100 Smaller volumes than IM; may be painful

(SC)

Oral (PO) 5 to < 100 Most convenient; first pass effects may be
significant

Rectal (PR) 30 to < 100 Less first-pass effects than oral

Inhalation 5 to < 100 Often very rapid onset

Transdermal 80 to ≤ 100 Usually very slow absorption; used for

lack of first-pass effects; prolonged duration
of action
 Objectives of bioavailability studies:

• Development of new formulation.

• Determination of influence of excipients, patient related
factors and possible interaction with other drugs on the
efficiency of absorption.
• Control of quality of a drug product during the early stages
of marketing in order to determine the influence of
processing factors, storage, stability on drug absorption.
• Primary stages of the development of a suitable dosage
form for a new drug entity.
 12
Plasma Concentration

10 TOXIC RANGE

6 THERAPEUTIC RANGE

2 SUB-THERAPEUTIC

0
0 1 2 3 4 5 6 7 8 9
Dose
DISPOSITION OF DRUGS
 ABSOLUTE BIOAVAILABILITY:
• The systemic availability of a drug administered orally is determined
in comparison to its iv administration.
• Characterization of a drug's absorption properties from the e.v. site.
• F = AUCev
AUCiv
RELATIVE BIOAVAILABILITY:
• The availability of a drug product as compared to another dosage
form or product of the same drug given in the same dose.
• Characterization of absorption of a drug from its formulation.
• Fr=AUCA
AUCB
 FACTORS INFLUENCING BIOAVAILABILITY:
• Three distinct factors are involved to influencing bioavailability.
These are:
1.Pharmaceutical factors:
• physicochemical properties of the drug.
1. Particle size
2. Crystalline structure
3. Salt form
• Formulation and manufacturing variables.
1.Disintegration and dissolution time
2.Pharmaceutical ingredients
3.Special coatings
4.Nature and type of dosage form
 2. Patient related factors:
• Physiologic factors.
1.Variations in pH of GI fluids
2.Gastric emptying rate
3. Intestinal motility
4. Presystemic and first-pass metabolism
5. Age, sex
6. Disease states
• Interactions with other substances.
1. Food
2. Fluid volume
3. Other drugs
3. Route of administration:
1.Parentral administration
2.Oral administration
3.Rectal administration
4.Topical administration
ASSESSMENT OF BIOAVAILABILITY

For assessing the bioavailability or clinical availability of a drug, its rate and
extent of absorption and its first-pass metabolism must be evaluated.

These criteria are difficult or even impossible to quantify.

The method used to assess bioavailability depend upon the assumption that
measurement of the drug concentration in a suitable body fluid, such as blood,
plasma, serum, urine or sometime saliva, can be correlated with its clinical
availability.

Bioavailability is determined by following methods

Pharmacokinetics method – This method is more practical and discriminative.

Pharmacokinetic methods are of two types.
a)Determination of whole blood, plasma or serum concentration
b)Urinary excretion method
 METHODS OF ASSESSING BIOAVAILABILITY:
• Bioavailability testing is a means of predicting the clinical efficacy of a
drug.

1.In-vivo methods:

1. Demonstration of a clinically significant effect.

• The drawbacks are complex, expensive, time-consuming and require
a sensitive and quantitative measure of the desired response.
2. Quantification of pharmacologic effect.
• This method is based on the assumption that a given intensity of
response is associated with a particular drug concentration at the site
of action.
• The drawbacks are monitoring of pharmacologic data is often
difficult, precision and reproducibility are difficult to establish.
• And there are only a limited number of pharmacologic effects (e.g.
heart rate, body temperature, blood sugar levels) that are applicable
to this method.
 2. Blood level studies:
• These are based on the assumption that there is a direct relationship between the
concentration of drug in blood or plasma and the concentration of drug at the site
of action.

Dosing interval 7
• The key parameters for determining bioavailability

1. AUC: The AUC is proportional to the total amount of

drug reaching the systemic circulation, and thus
characterizes the extent of absorption.

2. Cmax: Gives indication whether drug is sufficiently

absorbed systemically to provide a therapeutic
response.

3. Tmax: The Tmax reflects the rate of drug absorption,

and decreases as the absorption rate increases.
 3.Urinary Excretion Data:
• These studies are based on the premise that urinary excretion
of the unchanged drug is directly proportional to the plasma
concentration of total drug.

• This technique of studying bioavailability is most useful for

those drugs that are not extensively metabolized prior to
urinary elimination.
• The three major parameters examined in urinary excretion data
are as follow:
1.(dXu/dt)max : It gives the rate of appearance of drug in the urine is
proportional to its concentration in systemic circulation. Its value
increases as the rate of and/or extent of absorption increases

2. (tu)max : It is analogous to the of plasma level data, its value

decreases as the absorption rate increases.

3. Xu : It is related to the AUC of plasma level data and increases as

the extent of absorption increases.

• This studies used for certain thiazide diuretics and sulfonamides

and for drugs that have urine as the site of action-for example,
urinary antiseptics.
 DESIGN AND CONDUCT OF STUDIES:
• The design and conduct of the study should follow ICH regulations
on Good Clinical Practice, including reference to an Ethics
Committee.

• A bioequivalence study is basically a comparative bioavailability

study designed to establish equivalence between test and
reference products.

1. Design:
• The study should be designed in such a way that the formulation
effect can be distinguished from other effects.
• For the number of formulations the design of choice
is a two-period, two sequence crossover design.
• Parallel design, single dose studies, steady-state.
2. Subjects:
A.Selection of subjects:
• Aim to minimize variability and permit detection of differences
between pharmaceutical products.
• The studies should normally be performed with healthy
volunteers.
• They should be screened for suitability by means of clinical
laboratory tests, review of medical history, and medical
examination.
B.Standardization of the study:
• The test conditions should be standardized in order to minimize
the variability of all factors involved.
• Standardization of the diet, fluid intake and exercise is
recommended.
• The subjects should not take other medicines during a suitable
period before and during the study.
C.Inclusion of patients:
• If the investigated active substance is known to have adverse effects
and the pharmacological effects or risks are considered unacceptable
for healthy volunteers it may be necessary to use patients instead,
under suitable precautions and supervision.
D.Genetic phenotyping:
• Phenotyping of subjects should be considered as well in crossover
studies.
• If a drug is known to be subject to major genetic polymorphism,
studies could be performed in panels of subjects of known
phenotype or genotype.
3. Characteristics to be investigated:
• In most cases evaluation of bioavailability and bioequivalence will be
based upon the measured concentrations of the parent compound.
• In some situations, measurements of an active or inactive metabolite
is carried out.
• The plasma concentration versus time curves are mostly used to
assess extent and rate of absorption.
• The use of urine excretion data may be advantageous in determining
the extent of drug input in case of products predominately excreted
renally.
• Specificity, accuracy and reproducibility of the methods should be
sufficient.
4.Chemical analysis:
• It is conducted according to the applicable principles of Good
Laboratory Practice (GLP).
• Determination of the active moiety and/or its biotransformation
product(s) in plasma, urine or any other suitable matrix must be well
characterized, fully validated and documented to yield reliable results
that can be satisfactorily interpreted.
5 Reference and test product:
• The choice of reference product should be justified by the applicant
and agreed upon by the regulatory authority.
• The test products used in the biostudy must be prepared in
accordance with GMP-regulations.
6.Data analysis:
• To quantify the difference in bioavailability between the reference
and test products and to demonstrate that any clinically important
difference.
A.Statistical analysis:
• The statistical analysis (e.g. ANOVA) should take into account
sources of variation that can be reasonably assumed to have an
effect on the response variable.
• Pharmacokinetic parameters derived from measures of
concentration, e.g. AUC, Cmax should be analyzed using ANOVA.
B.Acceptance range for pharmacokinetic parameters:
1.AUC-ratio:
• It should lie within an acceptance interval of 0.80-1.25.
2.Cmax-ratio:
• It should lie within an acceptance interval of 0.80-1.25.
• The wider interval must be 0.75-1.33.
 For tmax if there is a clinically relevant claim for rapid release or
action or signs related to adverse effects. The interval should lie
within a clinically determined range.
C.Handling deviations from the study plan:
• The protocol should also specify methods for handling drop-outs.
D.A remark on individual and population bioequivalence:
• Bioequivalence studies are designed for population and individual
is limited.
7.In vitro dissolution for bioequivalence study:
• The term commonly used to describe is "in-vitro/in-vivo
correlation".
• The specifications for the in vitro dissolution of the product should
be derived from the dissolution profile of the batch.
A. Official dissolution tests:
• Apparatus 1, (basket method).
• Apparatus 2 (paddle method).
B.Parameters used:
1. Degree of agitation
2. Size and shape of container
3. Composition of dissolution medium
• pH, ionic strength, viscosity
4. Temperature of dissolution medium
5. Volume of dissolution medium
8.Reporting of results:
• The report of a bioequivalence study should give the complete
documentation of its protocol, conduct and evaluation
complying with GCP-rules and ICH guideline.
• Drop-out and withdrawal of subjects should be fully
documented.
• The method used to derive the pharmacokinetic parameters
should be specified.
• The analytical report should include the results for all standard
and quality control samples.
 APPLICATIONS FOR PRODUCTS CONTAINING NEW ACTIVE
SUBSTANCES:
1 Bioavailability:
• In the case of new active substances, the systemic availability of the
substance in its intended pharmaceutical form is measured in
comparison with intravenous administration.
2.Bioequivalence:
• Such studies may be exempted if the absence of differences in the in
vivo performance can be justified by satisfactory in vitro data.
APPLICATIONS FOR PRODUCTS CONTAINING APPROVED ACTIVE
SUBSTANCES:
A. Bioequivalence studies:
• In vivo bioequivalence studies are needed when there is a risk that
possible differences in bioavailability may result in therapeutic in
equivalence.
 • Bioequivalence (BE): Definition

“the absence of a significant difference in the rate and extent to

which the active ingredient or active moiety in pharmaceutical
equivalents or pharmaceutical alternatives becomes available at
the site of drug action when administered at the same molar dose
under similar conditions in an appropriately designed study.”
DEFINITION

Equivalence – Equivalence is more relative term that compares one

drug product with another or with a set of established standards.
Equivalence may be defined in several ways:

Chemical equivalence indicates that two or more dosage forms

contain the labelled quantities of drug.

Clinical equivalence occurs when the same drug from two or more
dosage forms gives identical in vivo effects as measured by a
pharmacological response or by control of a symptom or a disease.

Therapeutic equivalence implies that one structurally different

chemical can yield the same clinical result as another chemical.

Bioequivalence indicates that drug in two or more similar

dosage forms reaches the general circulation at the same
relative rate and the same relative extent.
• Bioequivalence
90
80
Concentration (ng/mL)

70
60
Tes t/Generic
50
Reference/B rand
40
30
20
10
0
0 5 10 15 20 25 30
Time (hours)
• Goals of BE

Ultimate: Bioequivalence studies impact of changes to

the dosage form process after pivotal studies commence
to ensure product on the market is comparable to
that upon which the efficacy is based

• Establish that a new formulation has therapeutic equivalence in the rate and
extent of absorption to the reference drug product.
• Important for linking the commercial drug product to clinical trial material at
time of NDA
• Important for post-approval changes in the marketed drug formulation
• Scheme of Oral Dosage Form

Human Intestinal
Absorption (HIA)

1,2 – Stability + Solubility

3 – Passive + Active Tr.
4 – Pgp efflux + CYP 3A4

Oral Bioavailability
(%F)
Why do we need Bioequivalence studies?

 No clinical studies have been performed in

patients with the Generic Product to support
its Efficacy and Safety.
 With data to support similar in vivo
performance (= Bioequivalence)
Efficacy and Safety
data can be extrapolated from the Innovator
Product to the Generic Product.
NEED FOR BIOEQUIVALENCE

•Bioequivalence studies provide a link between the

pivotal and early clinical trial formulation.

•Bioequivalence studies are for determination of the

therapeutic equivalence between the pharmaceutically
equivalencegeneric drug product and a corresponding
reference listed drug.

•Bioequivalence studies provide information on product

quality and performance when there are changes in
components, compositionand method of manufacture
after approval of the drug product.
Most common error made in bioavailability data is that of
cross – Study comparison
This occurs when the blood concentration – time curve of a drug
product in one study is compare with the blood concentration -
time curve of that drug product in another study.

There are three reasons

Different Subject Population:

Different Study Condition:

DIFFERENT ASSAY METHODOLOGY

 •FDA Requirements for
Bioequivalence
• Product A is bioequivalent
to the reference drug; its
125% 90% confidence interval of
Reference Range

the AUC falls within 80% to

Pharmacokinetic

125% of the reference drug

100% • Product B is not
bioequivalent to the
80% reference drug; its 90%
confidence interval of the
Product A
Bioequivalent
Reference
Drug
Product B
Not Bioequivalent
AUC falls outside of 80% to
125% of the reference drug

Approved Drug Products With Therapeutic Equivalence Evaluations. 23rd ed. 2003. FDA/CDER Web site.
Available at: http://www.fda.gov/cder/ob/docs/preface/ecpreface.htm#Therapeutic Equivalence-Related
Terms. Accessed September 29, 2003.
 LIMITATION OF BIOAVAILABILITY AND BIOEQUIVALENCE
A cross over design may be difficult for drugs with a long elimination half life.

Highly variable drugs may require a far greater number of subjects to meet the FDA

bioequivalence characteristics.

Certain characteristics in the biotransformation of drugs make it difficult to evaluate

the bioequivalence of such drugs. For e.g. for drugs that are stereoisomer with a

different rate of biotransformation and a different pharmacodynamic response, the

measurement of individual isomers may be difficult for analytical reasons.

Drugs that are administered by routes other than the oral route drugs/dosage forms

that are intended for local effects have minimal systemic bioavailability.
E.g. ophthalmic, dermal, intranasal and inhalation drug products.